Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1989 Jul 1;109(1):7–15. doi: 10.1083/jcb.109.1.7

Heat shock resistance conferred by expression of the human HSP27 gene in rodent cells

PMCID: PMC2115456  PMID: 2745558

Abstract

Heat shock induces in cells the synthesis of specific proteins called heat shock proteins (HSPs) and a transient state of thermotolerance. The putative role of one of the HSPs, HSP27, as a protective molecule during thermal stress has been directly assessed by measuring the resistance to hyperthermia of Chinese hamster and mouse cells transfected with the human HSP27 gene contained in plasmid pHS2711. One- and two-dimensional gel electrophoresis of [3H]leucine- and [32P]orthophosphate-labeled proteins, coupled with immunological analysis using Ha27Ab and Hu27Ab, two rabbit antisera that specifically recognize the hamster and the human HSP27 protein respectively, were used to monitor expression and inducibility of the transfected and endogenous proteins. The human HSP27 gene cloned in pHS2711 is constitutively expressed in rodent cells, resulting in accumulation of the human HSP27 and all phosphorylated derivatives. No modification of the basal or heat-induced expression of endogenous HSPs is detected. The presence of additional HSP27 protein provides immediate protection against heat shock administered 48 h after transfection and confers a permanent thermoresistant phenotype to stable transfectant Chinese hamster and mouse cell lines. Mild heat treatment of the transfected cells results in an induction of the full complement of the endogenous heat shock proteins and a small increase in thermoresistance, but the level attained did not surpass that of heat-induced thermotolerant control cells. These results indicate that elevated levels of HSP27 is sufficient to give protection from thermal killing. It is concluded that HSP27 plays a major role in the increased thermal resistance acquired by cells after exposure to HSP inducers.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arrigo A. P., Welch W. J. Characterization and purification of the small 28,000-dalton mammalian heat shock protein. J Biol Chem. 1987 Nov 15;262(32):15359–15369. [PubMed] [Google Scholar]
  2. Berger E. M., Woodward M. P. Small heat shock proteins in Drosophila may confer thermal tolerance. Exp Cell Res. 1983 Sep;147(2):437–442. doi: 10.1016/0014-4827(83)90225-2. [DOI] [PubMed] [Google Scholar]
  3. Chandler V. L., Maler B. A., Yamamoto K. R. DNA sequences bound specifically by glucocorticoid receptor in vitro render a heterologous promoter hormone responsive in vivo. Cell. 1983 Jun;33(2):489–499. doi: 10.1016/0092-8674(83)90430-0. [DOI] [PubMed] [Google Scholar]
  4. Chrétien P., Landry J. Enhanced constitutive expression of the 27-kDa heat shock proteins in heat-resistant variants from Chinese hamster cells. J Cell Physiol. 1988 Oct;137(1):157–166. doi: 10.1002/jcp.1041370119. [DOI] [PubMed] [Google Scholar]
  5. Craig E. A., Jacobsen K. Mutations of the heat inducible 70 kilodalton genes of yeast confer temperature sensitive growth. Cell. 1984 Oct;38(3):841–849. doi: 10.1016/0092-8674(84)90279-4. [DOI] [PubMed] [Google Scholar]
  6. Craig E. A. The heat shock response. CRC Crit Rev Biochem. 1985;18(3):239–280. doi: 10.3109/10409238509085135. [DOI] [PubMed] [Google Scholar]
  7. Gerner E. W., Schneider M. J. Induced thermal resistance in HeLa cells. Nature. 1975 Aug 7;256(5517):500–502. doi: 10.1038/256500a0. [DOI] [PubMed] [Google Scholar]
  8. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  9. Henle K. J., Dethlefsen L. A. Heat fractionation and thermotolerance: a review. Cancer Res. 1978 Jul;38(7):1843–1851. [PubMed] [Google Scholar]
  10. Hickey E., Brandon S. E., Potter R., Stein G., Stein J., Weber L. A. Sequence and organization of genes encoding the human 27 kDa heat shock protein. Nucleic Acids Res. 1986 May 27;14(10):4127–4145. doi: 10.1093/nar/14.10.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoffman E. P., Gerring S. L., Corces V. G. The ovarian, ecdysterone, and heat-shock-responsive promoters of the Drosophila melanogaster hsp27 gene react very differently to perturbations of DNA sequence. Mol Cell Biol. 1987 Mar;7(3):973–981. doi: 10.1128/mcb.7.3.973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ingolia T. D., Craig E. A. Four small Drosophila heat shock proteins are related to each other and to mammalian alpha-crystallin. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2360–2364. doi: 10.1073/pnas.79.7.2360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnston R. N., Kucey B. L. Competitive inhibition of hsp70 gene expression causes thermosensitivity. Science. 1988 Dec 16;242(4885):1551–1554. doi: 10.1126/science.3201244. [DOI] [PubMed] [Google Scholar]
  14. Kim Y. J., Shuman J., Sette M., Przybyla A. Nuclear localization and phosphorylation of three 25-kilodalton rat stress proteins. Mol Cell Biol. 1984 Mar;4(3):468–474. doi: 10.1128/mcb.4.3.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Landry J., Bernier D., Chrétien P., Nicole L. M., Tanguay R. M., Marceau N. Synthesis and degradation of heat shock proteins during development and decay of thermotolerance. Cancer Res. 1982 Jun;42(6):2457–2461. [PubMed] [Google Scholar]
  17. Landry J., Crête P., Lamarche S., Chrétien P. Activation of Ca2+-dependent processes during heat shock: role in cell thermoresistance. Radiat Res. 1988 Mar;113(3):426–436. [PubMed] [Google Scholar]
  18. Landry J., Samson S., Chrétien P. Hyperthermia-induced cell death, thermotolerance, and heat shock proteins in normal, respiration-deficient, and glycolysis-deficient Chinese hamster cells. Cancer Res. 1986 Jan;46(1):324–327. [PubMed] [Google Scholar]
  19. Lewis M. J., Pelham H. R. Involvement of ATP in the nuclear and nucleolar functions of the 70 kd heat shock protein. EMBO J. 1985 Dec 1;4(12):3137–3143. doi: 10.1002/j.1460-2075.1985.tb04056.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Li G. C., Werb Z. Correlation between synthesis of heat shock proteins and development of thermotolerance in Chinese hamster fibroblasts. Proc Natl Acad Sci U S A. 1982 May;79(10):3218–3222. doi: 10.1073/pnas.79.10.3218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lindquist S. The heat-shock response. Annu Rev Biochem. 1986;55:1151–1191. doi: 10.1146/annurev.bi.55.070186.005443. [DOI] [PubMed] [Google Scholar]
  22. Loomis W. F., Wheeler S. A. Chromatin-associated heat shock proteins of Dictyostelium. Dev Biol. 1982 Apr;90(2):412–418. doi: 10.1016/0012-1606(82)90390-6. [DOI] [PubMed] [Google Scholar]
  23. McAlister L., Finkelstein D. B. Heat shock proteins and thermal resistance in yeast. Biochem Biophys Res Commun. 1980 Apr 14;93(3):819–824. doi: 10.1016/0006-291x(80)91150-x. [DOI] [PubMed] [Google Scholar]
  24. Mestril R., Schiller P., Amin J., Klapper H., Ananthan J., Voellmy R. Heat shock and ecdysterone activation of the Drosophila melanogaster hsp23 gene; a sequence element implied in developmental regulation. EMBO J. 1986 Jul;5(7):1667–1673. doi: 10.1002/j.1460-2075.1986.tb04410.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  26. Pelham H. R. Hsp70 accelerates the recovery of nucleolar morphology after heat shock. EMBO J. 1984 Dec 20;3(13):3095–3100. doi: 10.1002/j.1460-2075.1984.tb02264.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pelham H. R. Speculations on the functions of the major heat shock and glucose-regulated proteins. Cell. 1986 Sep 26;46(7):959–961. doi: 10.1016/0092-8674(86)90693-8. [DOI] [PubMed] [Google Scholar]
  28. Petko L., Lindquist S. Hsp26 is not required for growth at high temperatures, nor for thermotolerance, spore development, or germination. Cell. 1986 Jun 20;45(6):885–894. doi: 10.1016/0092-8674(86)90563-5. [DOI] [PubMed] [Google Scholar]
  29. Pouysségur J., Franchi A., Salomon J. C., Silvestre P. Isolation of a Chinese hamster fibroblast mutant defective in hexose transport and aerobic glycolysis: its use to dissect the malignant phenotype. Proc Natl Acad Sci U S A. 1980 May;77(5):2698–2701. doi: 10.1073/pnas.77.5.2698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Regazzi R., Eppenberger U., Fabbro D. The 27,000 daltons stress proteins are phosphorylated by protein kinase C during the tumor promoter-mediated growth inhibition of human mammary carcinoma cells. Biochem Biophys Res Commun. 1988 Apr 15;152(1):62–68. doi: 10.1016/s0006-291x(88)80680-6. [DOI] [PubMed] [Google Scholar]
  31. Riabowol K. T., Mizzen L. A., Welch W. J. Heat shock is lethal to fibroblasts microinjected with antibodies against hsp70. Science. 1988 Oct 21;242(4877):433–436. doi: 10.1126/science.3175665. [DOI] [PubMed] [Google Scholar]
  32. Subjeck J. R., Shyy T. T. Stress protein systems of mammalian cells. Am J Physiol. 1986 Jan;250(1 Pt 1):C1–17. doi: 10.1152/ajpcell.1986.250.1.C1. [DOI] [PubMed] [Google Scholar]
  33. Thomas S. R., Lengyel J. A. Ecdysteroid-regulated heat-shock gene expression during Drosophila melanogaster development. Dev Biol. 1986 Jun;115(2):434–438. doi: 10.1016/0012-1606(86)90263-0. [DOI] [PubMed] [Google Scholar]
  34. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Voellmy R., Ahmed A., Schiller P., Bromley P., Rungger D. Isolation and functional analysis of a human 70,000-dalton heat shock protein gene segment. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4949–4953. doi: 10.1073/pnas.82.15.4949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Welch W. J. Phorbol ester, calcium ionophore, or serum added to quiescent rat embryo fibroblast cells all result in the elevated phosphorylation of two 28,000-dalton mammalian stress proteins. J Biol Chem. 1985 Mar 10;260(5):3058–3062. [PubMed] [Google Scholar]
  37. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wistow G., Piatigorsky J. Recruitment of enzymes as lens structural proteins. Science. 1987 Jun 19;236(4808):1554–1556. doi: 10.1126/science.3589669. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES